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PLANT BIOLOGY
Use of DNA barcodes to identify flowering plants

W. John Kress ^*, , Kenneth J. Wurdack ^*, , Elizabeth A. Zimmer ^*, Lee A. Weigt , and Daniel H. Janzen 

^*Department of Botany and Laboratories of Analytical Biology, National Museum of Natural History, Smithsonian Institution, P.O. Box 37012, Washington, DC 20013-7012; and Department of Biology, University of Pennsylvania, Philadelphia, PA 19104

Contributed by Daniel H. Janzen, April 15, 2005

Methods for identifying species by using short orthologous DNA sequences, known as "DNA barcodes," have been proposed and initiated to facilitate biodiversity studies, identify juveniles, associate sexes, and enhance forensic analyses. The cytochrome c oxidase 1 sequence, which has been found to be widely applicable in animal barcoding, is not appropriate for most species of plants because of a much slower rate of cytochrome c oxidase 1 gene evolution in higher plants than in animals. We therefore propose the nuclear internal transcribed spacer region and the plastid trnH-psbA intergenic spacer as potentially usable DNA regions for applying barcoding to flowering plants. The internal transcribed spacer is the most commonly sequenced locus used in plant phylogenetic investigations at the species level and shows high levels of interspecific divergence. The trnH-psbA spacer, although short (450-bp), is the most variable plastid region in angiosperms and is easily amplified across a broad range of land plants. Comparison of the total plastid genomes of tobacco and deadly nightshade enhanced with trials on widely divergent angiosperm taxa, including closely related species in seven plant families and a group of species sampled from a local flora encompassing 50 plant families (for a total of 99 species, 80 genera, and 53 families), suggest that the sequences in this pair of loci have the potential to discriminate among the largest number of plant species for barcoding purposes.

angiosperm | internal transcribed spacer | Plummers Island | species identification | trnH-psbA

Freely available online through the PNAS open access option.

Abbreviations: ITS, internal transcribed spacer; CO1, cytochrome c oxidase 1.

Data deposition: The sequences reported in this paper have been deposited in the GenBank database (accession nos. DQ005959-DQ006232).

To whom correspondence should be addressed. E-mail: kressj{at}si.edu.

© 2005 by The National Academy of Sciences of the USA

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